High level production of p-hydroxybenzoic acid in green plants

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/82 (2006.01) A01H 5/00 (2006.01) C12N 9/88 (2006.01) C12N 15/60 (2006.01) C12N 15/62 (2006.01)

Patent

CA 2411541

The invention relates to high-level production of pHBA in green plants using a unique expression cassette. The latter comprises a chorismate pyruvate lyase (CPL) coding sequence operably linked to a suitable promoter capable of driving protein expression in higher plants. Additionally, the CPL cassette comprises a sequence encoding a chloroplast transit peptide, its natural cleavage site, and a small portion of the transit peptide donor protein fused to the N-terminus of CPL. The chloroplast targeting sequence targets the foreign protein to the chloroplast compartment and aids in its uptake into the organelle. The cleavage site is unique to the transit peptide, and cleavage of the chimeric protein encoded by the cassette at this site releases a novel polypeptide that has full enzyme activity, comprising the mature CPL enzyme and a small portion of the transit peptide donor.

L'invention concerne une production de quantités élevées d'acide p-hydroxybenzoïque dans des plantes vertes au moyen d'une seule cassette d'expression. Cette dernière comprend une séquence de codage de Chorismate Pyruvate-Lyase (CPL) liée de manière fonctionnelle à un promoteur approprié pouvant entraîner l'expression de protéine dans des plantes supérieures. De plus, la cassette de CPL comprend une séquence codant un peptide de transit vers le chloroplaste, son site de restriction naturel, et une faible partie de la protéine donneuse du peptide de transit fusionnée avec l'extrémité N-terminale de CPL. La séquence de visée du chloroplaste oriente la protéine étrangère vers le compartiment du chloroplaste et facilite son absorption dans l'organelle. Le site de restriction est unique pour le peptide de transit, et une restriction de la protéine chimérique codée par la cassette à ce site délivre un nouveau polypeptide qui a une activité enzymatique complète, comprenant la CPL mature et une faible partie du donneur de peptide de transit.

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